A Molecule Printing Device for the Analysis of the Secretome of Single Cells

Abstract

The present invention discloses a device and methods for the analysis of secreted molecules from a single cell. As described herein, the invention incorporates individual microwells with a bottom surface capable of capturing a cell and allows the release of secreted molecules to be printed onto a capture surface. The device provides accurate identification of the cell source of the printed molecules by mapping the printed molecules to the cell source. The invention further employs spectrometry, immunoassay or label free Surface Plasmon Resonance imaging for detection of the secreted molecules in combination with a microwell array, where single cells are seeded in individual microwells and the secreted molecules are captured by the capture surface in an array print while the cell remains in the microwell for additional interrogation. The present invention has applications in medical research and diagnosis where individual target cells in a fluid sample are interrogated for the secreted products.

1 - A molecule printing device comprising:
 a. a microwell plate having individual microwells each with a bottom plate wherein at least one bottom plate has at least one precisely etched pore to pass a sample liquid containing molecules from a supply side to a discharge side; and   b. a capturing surface which is connected to the microwell plate such that molecules present in the sample liquid will move from the supply side of the pore towards the discharge side and captured on the capturing surface.   
     
     
         2 - The device according to  claim 1 , wherein the sample liquid contains components of interest with a slightly larger diameter than the pore such that when the sample fluid is applied to the microwell the object of interest will occlude the pore. 
     
     
         3 - The device according to  claim 2 , wherein the components of interest is a cell capable of partly or completely blocking the pore. 
     
     
         4 - The device according to  claim 2 , further having a means for retrieving captured components of interest. 
     
     
         5 - The device according to  claim 2 , wherein the molecules are secreted by the components of interest. 
     
     
         6 - The device of  claim 4  wherein the retrieving means is by a punching means of the bottom plate or a cell picking means using a micropipette. 
     
     
         7 - The device of  claim 1  having a capturing surface composition of any known surface for capturing molecules 
     
     
         8 - The device of  claim 7  having a capturing surface composition selected from the group consisting of glass, metal, ceramic, non-ceramic, organic, non-organic, nitrocellulose, paper, PVDF and combinations thereof. 
     
     
         9 - The device of  claim 7 , wherein the capturing surface contains capturing molecules that are able to capture the molecules present in the liquid. 
     
     
         10 - The device of  claim 9  having capturing molecules selected from the group consisting of antibodies, antigens, DNA, RNA, biotin, streptavidin and combinations thereof. 
     
     
         11 - The device of  claim 9  wherein the capturing surface is covered with a pattern of capturing molecules for a single microwell. 
     
     
         12 - The device of  claim 7  wherein the location of the microwells can be correlated to the printed molecules on the surface. 
     
     
         13 - The device of  claim 5  having a coupling means for maintaining a sterile and pressurized enclosure on the capturing surface while monitoring secreted molecules by surface plasmon resonance imaging (SPRi) comprising:
 a. a cup containing a microwell plate; and 
 b. a cap to maintain a pressurized and sterile enclosure. 
 
     
     
         14 - The device of  claim 13  wherein the pressure in the enclosure is determined by a thickness in the cap. 
     
     
         15 - The device of  claim 13  where the cup is bound to the microwell using tape. 
     
     
         16 - The device of  claim 15  wherein the tape is PEEK. 
     
     
         17 - The device of  claim 13  further having a means for connecting the microwell to the capture surface during transfer of molecules from the microwell to the capturing surface as a printed molecule, said means characterized by controlling a distance between the microwell and the capture surface. 
     
     
         18 - The device of  claim 17  wherein the distance is controlled by applying pressure to the top of the cap.